Abstract: To assess the stress action of cattle to handling, and then to provide reference to progress cattle handling, enrich the theory of transport stress of cattle, perfect the animal transport standard, body temperature and behavior of cattle in conditions of with and without chute in highway transport were measured. The results revealed that both of handlings with and without chute were stressful, with chute was more stressful than without chute, stress responses of cattle included the elevated temperature and behavior of escape, aggression, vocalization, and behavior strategy to resist handling stress were firstly escape, secondly aggression after escape were hindered and finally vocalization. The results of this study are valuable for evaluating and progressing animal handling mode.